Homework: Read ALL of Series 2:background information and all work performed in Labs 2-6 on our Classical (Forward) Genetics project. Write a summary of our experimental question & goals. Outline the experimental process, including all parts of the experimental design: Mutant Hunt, Linkage Analysis, Mapping the Mutation to a chromosome and to a particular location, Complementation Analysis, Determining whether or not the mutation has been previously characterized, & Defining the gene sequence and protein change through DNA sequence analysis. Use this outline to write a 1-2 page summary of how you will determine the exact location and extent of the gene and protein change and include the significance of the mutation in worms and, if possible, broader significance in other species. Assignment explained and rubric found at BISC_219/F10: Assignment Series2_Outline_Summary

Start Data Analysis (Results) of your autosomal vs. X-linked testing DUE at the beginning of LAB 4. Grading rubric & Assignment info at:BISC_219/F10: Assignment Help- Data Analysis 1; Read the journal article, Indentification of Genes that Regulate a Left-Right Aymmetric Neuronal Migration in Caenorhabditis elegans published in Genetics 164: 1355-1367 (August 2003), for discussion in lab next time. Full text of article found at: [1]; See BISC_219/F10:Questions to Guide Your Reading1 for information on how to prepare for this discussion

Homework: Draw crosses and diagram strategy for Mapping the Location of your dpy mutation. Due at the beginning of Lab 5. Assignment described at BISC_219/F10: Assignment_Series2_Mapping Crosses; Read the journal article by Davis at al.,Rapid single nucleotide polymorphism mapping in C. elegans, found at BMC Genomics 2005, 6:118 doi:10.1186/1471-2164-6-118 and compare their mapping strategy to yours.

Homework: Explain complementation analysis in general and specifically how it will be used to confirm and expand your characterization of your dpy mutation. Due at the beginning of Lab 6. Assignment described atBISC_219/F10: Assignment_Series2_Complementation

Day before lab: grow overnight culture of single colony from transformation

Homework: Write all Series3 Protocols as Materials & Methods. Assignment described at BISC_219/F10: Assignment_Series3_ Materials and Methods. Due at the beginning of Lab 9 ; Read the original 1998 paper by Fire and Mello published in Nature391: 806-811 (Feb.19 1998) at doi:10.1038/35888 ; Do your own library search to find and read a recent review article about RNAi in C. elegans. Also read the 2002 Simmer et al. paper in Current Biology12:1317-1319 explaining the rrf-3 strain that we will use for our RNAi work at doi:10.1016/S0960-9822(02)01041-2 . These articles provide background information on how the mechanism of RNAi was worked out experimentally and will be helpful to you when you write the introduction section of your Series3 paper. Also read a published journal article by Green et al. (2009) Impact of Cigarette Smoke Exposure on Innate Immunity: A Caenorhabditis elegans Model. PLoS ONE 4(8): e6860, found at doi:10.1371/journal.pone.0006860. This study, like yours, uses reverse genetics to investigate gene function. There are questions to guide your reading found at:BISC_219:Questions to Guide Your Reading 2 . Be prepared to discuss those questions in Lab 9.

Homework: Construct figures/tables with properly formatted legends to illustrate the main findings of the RNAi part of your Series3:Reverse Genetics project. These figures are a draft of those that will be part of the results section of the research report you will write for Series3. Due on Nov. 24 for all students.

NO LAB

Nov. 18 to Nov. 26

Thanksgiving Break

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11

Nov. 29 to Dec. 3

Series 3:Reverse Genetics RT PCR;

Instructor will run gel; when results available review and interpret gel image